体外诱导骨肉瘤特异性抗肿瘤的免疫实验

采用分离骨肉瘤患者外周血单核细胞体外诱导DC细胞,Trizol法提取患者骨肉瘤细胞总RNA,用总RNA转染DC并诱导特异性CTL的扩增,用MTT法检测淋巴细胞的增殖和CTL的杀伤活性。探讨骨肉瘤细胞总RNA转染的DC疫苗体外诱导特异性抗肿瘤免疫的能力。经骨肉瘤细胞总RNA转染的DC特异性表面标志及功能相关分子表达均上调,转染后的DC可显著刺激自体T淋巴细胞增殖,诱导的特异性CTL对靶细胞的杀伤率显著高于单纯淋巴细胞和未经转染的DC。说明骨肉瘤细胞总RNA转染的DC疫苗可在体外诱导出特异性抗肿瘤免疫。

Experimental Study of Specific Antitumor Immunity Induced by Dendritic Cell Vaccine Transfected with Tumor Cell Total RNA in Bone Sarcoma Patients

To observe ability of DC vaccine transfected with tumor cell total RNA to induce specific antitumor immunity in bone sarcoma patients in vitro,DCs were generated from bone sarcoma patients' peripheral blood mononuclear cells(PBMC).Total RNA was isolated from bone sarcoma cell by Trizol.Autologous DCs transfected with total RNA of bone sarcoma cell were used to induce specific CTL proliferation.T cells proliferation and specific cytotoxicity were measured by MTT method.Transfected DCs exhibited dramatically increased expression of specific membrane markers and function-associated molecules,and were more potent in stimulating autologous T cell proliferation than that of control DCs.Specific CTLs induced by transfeced DCs showed higher cytotoxicity than LAK and control DCS against sarcoma cells.It is conclued that bone sarcoma patients' autologous DCs transfected with tumor total RNA are effective vaccines in stimulating specific antitumor T-cell immunity in vitro.